Abstract

Reversed-phase high-performance liquid chromatography (RP-HPLC) has greatly influenced and stimulated research in detection, quantitation, and identification of normal and abnormal globin chains in human newborn and adult red blood cells. In fact, RP-HPLC methodology has a number of advantages: it is fast and accurate, minute amounts of material are sufficient for each determination, complete automation is possible, and, most importantly, identification of electrophoretically "silent" mutant
hemoglobins due to neutral-to-neutral amino acid substitutions is feasible and reliable. It also greatly facilitates the separation and isolation of even microquantities of a mutant globin chain, and of globin proteolytic fragments, for subsequent characterization of the substitution within the abnormal peptide.